New method to deliver exogenous material into developing planarian embryos.
The ability to report or modify the embryological processes in living embryos is pivotal for developmental biology research. Planarian embryology has experienced renewed interest as the genetic pathways that drive adult regeneration were found to be involved in the development of embryos. The major drawback to the study of planarian embryology is the absence of methods that give access to the embryos and enable their manipulation. Herein, we report on a novel method for delivering external material into developing embryos using nanosecond laser pulses. When focused on the eggshell surface under optimal parameters, laser pulses ablate the protective case and open a pathway throughout which foreign material can be delivered. In this study, we used egg capsules from Schmidtea polychroa (Schmidt, 1861) to microinject 1 microm fluorescein isothiocyanate fluorescent beads into the live embryos. We obtained viability values ranging from 15% in early egg capsules to 100% in late developmental stages. Moreover, we measured the delivery effectiveness as the number of hatchlings containing fluorescent beads per microinjected egg capsule, reaching 100% in early stages and almost 40% in late stages. This is the first time that planarian embryos have been modified without compromising normal development. We consider that this technique will be of extreme value to future work on planarian developmental biology and regeneration, enabling the application of modern functional tools to the study of this Lophotrochozoan.
Martín-Durán
José María
JM
Departament de Genètica, Universitat de Barcelona, Barcelona, Spain. chemamartin@ub.edu
Duocastella
Martí
M
Serra
Pere
P
Romero
Rafael
R
D000818:Animals; D004625:Embryo, Nonmammalian / Q000528:radiation effects; D016650:Fluorescein-5-isothiocyanate; D007834:Lasers; D010539:Permeability / Q000528:radiation effects; D010932:Planarians / Q000196:embryology / Q000502:physiology*; D014162:Transfection / Q000379:methods*; D015053:Zygote / Q000737:chemistry / Q000502:physiology / Q000648:ultrastructure
D016650:Fluorescein-5-isothiocyanate